Analysis of ergosterol in single kernel and ground grain by gas chromatography-mass spectrometry

Yanhong Dong, Brian J. Steffenson, Chester J. Mirocha

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

A method for analyzing ergosterol in a single kernel and ground barley and wheat was developed using gas chromatography-mass spectrometry (GC-MS). Samples were saponified in methanolic KOH. Ergosterol was extracted by "one step" hexane extraction and subsequently silylated by N- trimethylsilylimidazole/trimethylchlorosilane (TMSI/TMCS) reagent at room temperature. The recoveries of ergosterol from ground barley were 96.6, 97.1, 97.1, 88.5, and 90.3% at the levels of 0.2, 1, 5, 10, and 20 μg/g (ppm), respectively. The recoveries from a single kernel were between 93.0 and 95.9%. The precision (coefficient of variance) of the method was in the range 0.8-12.3%. The method detection limit (MDL) and the method quantification limit (MQL) were 18.5 and 55.6 ng/g (ppb), respectively. The ergosterol analysis method developed can be used to handle 80 samples daily by one person, making it suitable for screening cereal cultivars for resistance to fungal infection. The ability for detecting low levels of ergosterol in a single kernel provides a tool to investigate early fungal invasion and to study mechanisms of resistance to fungal diseases.

Original languageEnglish (US)
Pages (from-to)4121-4125
Number of pages5
JournalJournal of agricultural and food chemistry
Volume54
Issue number12
DOIs
StatePublished - Jun 14 2006

Keywords

  • Barley
  • Ergosterol
  • GC-MS
  • Single kernel
  • Wheat

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